Plasmid-mediated transfer of antibiotic resistance genes and biofilm formation in a simulated drinking water distribution system under chlorine pressure. - Related Documents




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719301.0000Plasmid-mediated transfer of antibiotic resistance genes and biofilm formation in a simulated drinking water distribution system under chlorine pressure. The effects of disinfectants and plasmid-based antibiotic resistance genes (ARGs) on the growth of microorganisms and the plasmid-mediated transfer of ARGs in the water and biofilm of the drinking water distribution system under simulated conditions were explored. The heterotrophic plate count of the water in reactors with 0.1 mg/L NaClO and NH(2)Cl was higher than in the control groups. There was no similar phenomenon in biofilm. In the water of reactors containing NaClO, the aphA and bla genes were lower than in the antibiotic resistant bacteria group, while both genes were higher in the water of reactors with NH(2)Cl than in the control group. Chloramine may promote the transfer of ARGs in the water phase. Both genes in the biofilm of the reactors containing chlorine were lower than the control group. Correlation analysis between ARGs and water quality parameters revealed that the copy numbers of the aphA gene were significantly positively correlated with the copy numbers of the bla gene in water and significantly negatively correlated in biofilm (p < 0.05). The results of the sequencing assay showed that bacteria in the biofilm, in the presence of disinfectant, were primarily Gram-negative. 1.0 mg/L chlorine decreased the diversity of the community in the biofilm. The relative abundance of some bacteria that may undergo transfer increased in the biofilm of the reactor containing 0.1 mg/L chlorine.202539617560
719410.9999Response of antibiotic resistance genes in constructed wetlands during treatment of livestock wastewater with different exogenous inducers: Antibiotic and antibiotic-resistant bacteria. This work aimed to study the behavior of antibiotic resistance genes (ARGs) in constructed wetlands with different exogenous inducers additions (oxytetracycline and its resistant bacteria) by high-throughput quantitative polymerase chain reaction. Results indicated that constructed wetlands have the potential to reduce ARGs relative abundances in wastewater, and the total ARGs removal efficiency could exceed 60%. ARGs profile in the effluent differed from that in the influent, and that did not directly reflect the export of dominant ARGs in wetland biofilms. Meanwhile, the highest levels of detected numbers and relative abundances of ARGs were 43 and 3.35 × 10(-1) for control system and 44 and 6.40 × 10(-1) for treatment system, respectively, which meant that ARGs generation in wetlands were inevitable, and antibiotic and antibiotic-resistant bacteria from wastewater could indeed promote ARGs abundance in the system. Compared to the single roles of inducers, their synergistic role had a more significant influence on ARGs relative abundance.202032652450
719520.9999Insight into effect of high-level cephalexin on fate and driver mechanism of antibiotics resistance genes in antibiotic wastewater treatment system. In the study, antibiotic resistance genes (ARGs) were examined in wastewater and sludge samples to explore the effect of cephalexin (CFX) on the spreading and removal of ARGs in the Expanded Granular Sludge Bed (EGSB) reactor treating antibiotics wastewater. The result showed that the addition of CFX in the wastewater affected the removal amount of β-lactam ARGs and other types ARGs. Besides, the addition of CFX in the wastewater had no obviously effect on total concentration of targeted ARGs in the sludge, but it was related to the accumulation of some typical ARGs. Based on gene cassette array libraries analysis, the diversity of gene cassettes carried by intI1 gene was increased by the addition of CFX in the wastewater. Furthermore, the co-occurrence patterns between ARGs and bacterial genus were also investigated. The results showed the CFX in the wastewater not only affected the number of potential host bacteria of ARGs, but also changed the types of potential host bacteria of ARGs. The correlation analysis of ARG in influent, effluent and sludge showed that, for blaCTX-M, sul2, qnrS and AmpC genes, their removal amount in EGSB reactor treating antibiotic wastewater system might be enhanced by reducing their concentration in the sludge.202032505047
763030.9999Coexistence of silver ion and tetracycline at environmentally relevant concentrations greatly enhanced antibiotic resistance gene development in activated sludge bioreactor. Antibiotic resistance has become a global public health problem. Recently, various environmental pollutants have been reported to induce the proliferation of antibiotic resistance. However, the impact of multiple pollutants (e.g., heavy metals and antibiotics), which more frequently occur in practical environments, is poorly understood. Herein, one widely distributed heavy metal (Ag(+)) and one frequently detected antibiotic (tetracycline) were chosen to investigate their coexisting effect on the proliferation of antibiotic resistance in the activated sludge system. Results show that the co-occurrence of Ag(+) and tetracycline at environmentally relevant concentrations exhibited no distinct inhibition in reactor performances. However, they inhibited the respiratory activity by 42%, destroyed the membrane structure by 218%, and increased membrane permeability by 29% compared with the blank control bioreactor. Moreover, the relative abundances of target antibiotic resistance genes (ARGs) (e.g., tetA, bla(TEM-1), and sulII) in effluent after exposure of coexisting Ag(+) and tetracycline were increased by 92-1983% compared with those in control reactor, which were 1.1-4.3 folds higher than the sum of the sole ones. These were possibly attributed to the enrichments of antibiotic-resistant bacteria. The results would illumine the coexisting effect of heavy metals and antibiotics on the dissemination of ARGs in activated sludge system.202234482077
719040.9999Dynamics of microbial community and tetracycline resistance genes in biological nutrient removal process. The occurrence of antibiotics in wastewater has become a serious concern due to the possible development of antibiotic resistant bacteria in wastewater treatment process. In order to understand the dynamics of microbial community and tetracycline resistance genes in biological nutrient removal (BNR) process, three lab-scale sequencing batch reactors (SBRs) were operated under the stress of tetracycline. Results indicated that microbial community structure was altered, and tetracycline efflux pump genes were enhanced over 150-day operation in the presence of trace tetracycline of 20 and 50 μg L(-1), respectively. Furthermore, when the initial tetracycline concentrations were increased to 2 and 5 mg L(-1), substantial enhancement of tetracycline resistance was observed, accompanied with a sharp shift in microbial community structure. In this study, horizontal gene transfer was found to be the main mechanism for the development of tetracycline resistance genes under the long-terms stress of trace tetracycline. About 90.34% of the observed variations in tetracycline resistance genes could be explained by the dynamics of potential hosts of tetracycline resistance genes and class 1 integron. It should be noticed that the functional bacteria (e.g. Nitrospira, Dechloromonas, Rhodobacter and Candidatus_Accumulibacter) responsible for nutrient removal were positively correlated with tetracycline resistance, which might promote the prevalence of tetracycline resistance during biological wastewater treatment. Consequently, this study provided in-depth insights into the occurrence and prevalence of tetracycline resistance genes and their microbial hosts in BNR process.201930849601
719150.9999The impact of antimicrobials on the efficiency of methane fermentation of sewage sludge, changes in microbial biodiversity and the spread of antibiotic resistance. The study was designed to simultaneously evaluate the influence of high doses (512-1024 µg/g) the most commonly prescribed antimicrobials on the efficiency of anaerobic digestion of sewage sludge, qualitative and quantitative changes in microbial consortia responsible for the fermentation process, the presence of methanogenic microorganisms, and the fate of antibiotic resistance genes (ARGs). The efficiency of antibiotic degradation during anaerobic treatment was also determined. Metronidazole, amoxicillin and ciprofloxacin exerted the greatest effect on methane fermentation by decreasing its efficiency. Metronidazole, amoxicillin, cefuroxime and sulfamethoxazole were degraded in 100%, whereas ciprofloxacin and nalidixic acid were least susceptible to degradation. The most extensive changes in the structure of digestate microbiota were observed in sewage sludge exposed to metronidazole, where a decrease in the percentage of bacteria of the phylum Bacteroidetes led to an increase in the proportions of bacteria of the phyla Firmicutes and Proteobacteria. The results of the analysis examining changes in the concentration of the functional methanogen gene (mcrA) did not reflect the actual efficiency of methane fermentation. In sewage sludge exposed to antimicrobials, a significant increase was noted in the concentrations of β-lactam, tetracycline and fluoroquinolone ARGs and integrase genes, but selective pressure was not specific to the corresponding ARGs.202133831706
719260.9999Sulfamethoxazole and COD increase abundance of sulfonamide resistance genes and change bacterial community structures within sequencing batch reactors. The abundant microbial community in biological treatment processes in wastewater treatment plants (WWTPs) may potentially enhance the horizontal gene transfer of antibiotic resistance genes with the presence of antibiotics. A lab-scale sequencing batch reactor was designed to investigate response of sulfonamide resistance genes (sulI, sulII) and bacterial communities to various concentrations of sulfamethoxazole (SMX) and chemical oxygen demand (COD) of wastewater. The SMX concentrations (0.001 mg/L, 0.1 mg/L and 10 mg/L) decreased with treatment time and higher SMX level was more difficult to remove. The presence of SMX also significantly reduced the removal efficiency of ammonia nitrogen, affecting the normal function of WWTPs. All three concentrations of SMX raised both sulI and sulII genes with higher concentrations exhibiting greater increases. The abundance of sul genes was positive correlated with treatment time and followed the second-order reaction kinetic model. Interestingly, these two genes have rather similar activity. SulI and sulII gene abundance also performed similar response to COD. Simpson index and Shannon-Weiner index did not show changes in the microbial community diversity. However, the 16S rRNA gene cloning and sequencing results showed the bacterial community structures varied during different stages. The results demonstrated that influent antibiotics into WWTPs may facilitate selection of ARGs and affect the wastewater conventional treatment as well as the bacteria community structures.201728211331
718270.9999Effects of UV disinfection on phenotypes and genotypes of antibiotic-resistant bacteria in secondary effluent from a municipal wastewater treatment plant. To elucidate the effects of UV disinfection on antibiotic resistance in biologically-treated wastewater, we investigated the antibiotic resistance profiles, species of cultivable heterotrophic bacteria, and antibiotic-resistance genes (ARGs) in antibiotic-resistant bacteria before and after treatment. UV disinfection greatly changed the bacterial community structure and the antibiotic resistance in wastewater. The antibiotic resistance in wastewater samples was strongly associated with the bacterial community. The proportions of Gram-positive bacteria gradually increased with increasing UV fluence. The proportions of bacteria resistant to cephalexin, penicillin, and vancomycin all greatly decreased after UV treatment in both sampling events (July 2018 and January 2019), and those for bacteria resistant to ofloxacin, ciprofloxacin, and sulfadiazine increased, resulting from the alternative antibiotic resistance profiles among different genera. UV disinfection induced the selection of multi-antibiotic resistant (MAR) bacteria. For example, the MAR indices of Aeromonas, the dominant genus during the treatments, were significantly increased after UV irradiation (P < 0.05). The MAR index was also markedly increased (P < 0.05) at a fluence of 5 mJ/cm(2) in both events. In UV10 treatment, the bacterial community structure was greatly changed. The genera with relatively low MAR indices replaced that with high MAR indices, and became the dominant genera. As a result, the MAR indices of treated samples showed a decreased trend after 10 mJ/cm(2) UV irradiation. The detection frequencies of ARGs located on the chromosome varied mainly due to the evolution of the microbial community. The occurrence of ARGs (tetA, tetC, tetM, tetW, tetX, and sul1) located on plasmid DNA decreased after UV disinfection, and the average detection frequencies of tet and sul genes decreased by 15% and 6%, respectively (P < 0.05). Generally speaking, the effect of UV disinfection on the enrichment of antibiotic resistance is limited in this study, and horizontal gene transfer via the plasmids in surviving bacteria might be impaired due to the decreased abundance of ARGs on the plasmids.201930991178
757180.9998Ancient Oriental Wisdom still Works: Removing ARGs in Drinking Water by Boiling as compared to Chlorination. Antibiotic resistance genes (ARGs) in municipal drinking water may not be effectively removed during centralized treatment. To reduce potential health risks, water disinfection at the point-of-use scale is warranted. This study investigated the performance of boiling, a prevalent household water disinfection means, in response to ARGs contamination. We found that boiling was more efficient in inactivating both Escherichia coli and environmental bacteria compared to chlorination and pasteurization. Boiling of environmental bacteria suspension removed a much broader spectrum of ARGs and mobile genetic elements (up to 141 genes) than chlorination (up to 13 genes), such better performance was largely attributed to a stronger inactivation of chlorine-tolerant bacteria including Acinetobacter and Bacillus. Accumulation of extracellular ARGs was found during low-temperature heating (≤ 80°C) and in the initial stage of chlorination (first 3 min when initial chlorine was 5 mg/L and first 12 min when initial chlorine was 1 mg/L). These extracellular ARGs as well as the intracellular ARGs got removed as the heating temperature increased or the chlorination time prolonged. Under the same treatment time (30 min), high-temperature heating (≥ 90.1°C) damaged the DNA structure more thoroughly than chlorination (5 mg/L). Taking into account the low transferability of ARGs after DNA melting, boiling may provide an effective point-of-use approach to attenuating bacterial ARGs in drinking water and is still worth promoting in the future.202234910990
780190.9998Disinfection of swine wastewater using chlorine, ultraviolet light and ozone. Veterinary antibiotics are widely used at concentrated animal feeding operations (CAFOs) to prevent disease and promote growth of livestock. However, the majority of antibiotics are excreted from animals in urine, feces, and manure. Consequently, the lagoons used to store these wastes can act as reservoirs of antibiotics and antibiotic-resistant bacteria. There is currently no regulation or control of these systems to prevent the spread of these bacteria and their genes for antibiotic resistance into other environments. This study was conducted to determine the disinfection potential of chlorine, ultraviolet light and ozone against swine lagoon bacteria. Results indicate that a chlorine dose of 30 mg/L could achieve a 2.2-3.4 log bacteria reduction in lagoon samples. However, increasing the dose of chlorine did not significantly enhance the disinfection activity due to the presence of chlorine-resistant bacteria. The chlorine resistant bacteria were identified to be closely related to Bacillus subtilis and Bacillus licheniformis. A significant percentage of lagoon bacteria were not susceptible to the four selected antibiotics: chlortetracycline, lincomycin, sulfamethazine and tetracycline (TET). However, the presence of both chlorine and TET could inactivate all bacteria in one lagoon sample. The disinfection potential of UV irradiation and ozone was also examined. Ultraviolet light was an effective bacterial disinfectant, but was unlikely to be economically viable due to its high energy requirements. At an ozone dose of 100 mg/L, the bacteria inactivation efficiency could reach 3.3-3.9 log.200616678233
8005100.9998Deciphering the fate of antibiotic resistance genes in norfloxacin wastewater treated by a bio-electro-Fenton system. The misuse of antibiotics has increased the prevalence of antibiotic resistance genes (ARGs), considered a class of critical environmental contaminants due to their ubiquitous and persistent nature. Previous studies reported the potentiality of bio-electro-Fenton processes for antibiotic removal and ARGs control. However, the production and fate of ARGs in bio-electro-Fenton processes triggered by microbial fuel cells are rare. In this study, the norfloxacin (NFLX) average residual concentrations within two days were 2.02, 6.07 and 14.84 mg/L, and the average removal efficiency of NFLX was 79.8 %, 69.6 % and 62.9 % at the initial antibiotic concentrations of 10, 20 and 40 mg/L, respectively. The most prevalent resistance gene type in all processes was the fluoroquinolone antibiotic gene. Furthermore, Proteobacteria was the dominant ARG-carrying bacteria. Overall, this study can provide theoretical support for the efficient treatment of high antibiotics-contained wastewater by bio-electro-Fenton systems to better control ARGs from the perspective of ecological security.202236252757
7189110.9998Comparative effects of different antibiotics on antibiotic resistance during swine manure composting. This study explored commonly-used antibiotics (lincomycin, chlorotetracycline, sulfamethoxazole, and ciprofloxacin) and their collective effects on antibiotic resistance during composting. In the first 7 days, ciprofloxacin showed the greatest influence on the physicochemical factors among the studied antibiotics; the removal of antibiotic resistance genes (ARGs) in the multiple-antibiotic treatment was significantly less than single-antibiotic treatments; especially, the largest removal of ribosomal protection genes (tetW and tetO) occurred in single ciprofloxacin treatment. In the end of composting, similar removal ratio (29.71-99.79%) of ARGs was achieved in different treatments (p greater than 0.05); Chloroflexi became the main phylum and it was closely associated with ARGs removal based on the network analysis. Potential host bacteria of ARGs varied with different antibiotics; in particular, the presence of multiple antibiotics increased potential host bacteria of ermA, sul1 and tetO. Above all, collective effects of different antibiotics led to the enrichment of antibiotic resistance in the composting.202032712514
7022120.9998Fate of antibiotics, antibiotic-resistant bacteria, and cell-free antibiotic-resistant genes in full-scale membrane bioreactor wastewater treatment plants. The removal of antibiotics, antibiotic-resistant bacteria (ARB), and cell-free antibiotic-resistant genes (ARGs) and the microbial community of ARB were investigated in detail to understand their fate and provide valuable information on the feasibility of full-scale membrane bioreactor (MBR). The potential risks of cell-free ARGs to the receiving environment were discovered. High influent antibiotic concentration could inhibit the microbial activity of MBR sludge, whereas good antibiotic removal could be maintained because of relatively long solid retention time and high biomass retention. Approximately 61.8%-77.5% of the total antibiotics were degraded, and 22.5%-38.2% of the total antibiotics were adsorbed by MBR sludge on average. The individual antibiotic removal presented intense discrepancy because of the chemical construction and distribution coefficient of antibiotics. Aeromonas exhibited specific antibiotic resistance to ampicillin and erythromycin, Escherichia became the predominant genera in kanamycin-ARB and tetracycline-ARB, and Klebsiella and Bacteroides were the particular genera that exhibited distinct antibiotic resistance to ciprofloxacin. A significant correlation was found between cell-free ARG abundance and ARB content, and relatively high effluent cell-free ARG abundance facilitated the proliferation and transmission of ARB. The impacts of the receiving environment to eliminate the ecological risks and severe threats to human health should be investigated because of the low decay ratio and long-term persistence of cell-free ARGs.202031986335
3432130.9998Insights into the amplification of bacterial resistance to erythromycin in activated sludge. Wastewater treatment plants are significant reservoirs for antimicrobial resistance. However, little is known about wastewater treatment effects on the variation of antibiotic resistance. The shifts of bacterial resistance to erythromycin, a macrolide widely used in human medicine, on a lab-scale activated sludge system fed with real wastewater was investigated from levels of bacteria, community and genes, in this study. The resistance variation of total heterotrophic bacteria was studied during the biological treatment process, based on culture dependent method. The alterations of bacterial community resistant to erythromycin and nine typical erythromycin resistance genes were explored with molecular approaches, including high-throughput sequencing and quantitative polymerase chain reaction. The results revealed that the total heterotrophs tolerance level to erythromycin concentrations (higher than 32 mg/L) was significantly amplified during the activated sludge treatment, with the prevalence increased from 9.6% to 21.8%. High-throughput sequencing results demonstrated an obvious increase of the total heterotrophic bacterial diversity resistant to erythromycin. Proteobacteria and Bacteroidetes were the two dominant phyla in the influent and effluent of the bioreactor. However, the prevalence of Proteobacteria decreased from 76% to 59% while the total phyla number increased greatly from 18 to 29 through activated sludge treatment. The gene proportions of erm(A), mef(E) and erm(D) were greatly amplified after biological treatment. It is proposed that the transfer of antibiotic resistance genes through the variable mixtures of bacteria in the activated sludge might be the reason for the antibiotic resistance amplification. The amplified risk of antibiotic resistance in wastewater treatment needs to be paid more attention.201525957255
7201140.9998Evolution of microbial community and drug resistance during enrichment of tetracycline-degrading bacteria. Recently, the microbial degradation of tetracycline has been widely reported. However, its potential risks in treating wastewater containing high concentrations of tetracycline have not been fully evaluated. In this study, the evolution of the microbial community and drug resistance was traced during the enrichment of tetracycline-degrading bacteria. The results showed that some minor compositions such as Shewanella, Bacillus, and Pseudomonas in the seed sludge became the predominant genera in the enrichment cultures when continuously using tetracycline as the sole carbon source, especially some possible pathogenic bacteria increased significantly in this process. The abundances of most TRGs/16S rDNA were increased after enrichment, although the relative abundance of tetA and tetL genes decreased to some extent. From the enrichment culture, 7 predominant tetracycline-degrading strains were isolated, of which TD-1 (Bacillus) and TD-5 (Shewanella) presented high degradation efficiencies (6-day degradation rate > 95%, half-life period of tetracycline ≈ 24 h). In addition, multiple TRGs, mobile genetic elements (MGEs) and even gene cassettes were found in each tetracycline-degrading isolate. The findings suggested that some risks such as the pathogenicity of isolates and the spread of ARGs should be considered when the biodegradation method is used to treat wastewater polluted with high concentrations of tetracycline.201930660087
7200150.9998Plant Growth, Antibiotic Uptake, and Prevalence of Antibiotic Resistance in an Endophytic System of Pakchoi under Antibiotic Exposure. Antibiotic contamination in agroecosystems may cause serious problems, such as the proliferation of various antibiotic resistant bacteria and the spreading of antibiotic resistance genes (ARGs) in the environment or even to human beings. However, it is unclear whether environmental antibiotics, antibiotic resistant bacteria, and ARGs can directly enter into, or occur in, the endophytic systems of plants exposed to pollutants. In this study, a hydroponic experiment exposing pakchoi (Brassica chinensis L.) to tetracycline, cephalexin, and sulfamethoxazole at 50% minimum inhibitory concentration (MIC) levels and MIC levels, respectively, was conducted to explore plant growth, antibiotic uptake, and the development of antibiotic resistance in endophytic systems. The three antibiotics promoted pakchoi growth at 50% MIC values. Target antibiotics at concentrations ranging from 6.9 to 48.1 µg·kg(-1) were detected in the treated vegetables. Additionally, the rates of antibiotic-resistant endophytic bacteria to total cultivable endophytic bacteria significantly increased as the antibiotics accumulated in the plants. The detection and quantification of ARGs indicated that four types, tetX, bla(CTX-M), and sul1 and sul2, which correspond to tetracycline, cephalexin, and sulfamethoxazole resistance, respectively, were present in the pakchoi endophytic system and increased with the antibiotic concentrations. The results highlight a potential risk of the development and spread of antibiotic resistance in vegetable endophytic systems.201729099753
7184160.9998Effects of activated sludge and UV disinfection processes on the bacterial community and antibiotic resistance profile in a municipal wastewater treatment plant. Wastewater tertiary treatment has been pointed out as an effective alternative for reducing the concentration of antibiotic resistant bacteria and genes (ARB and ARGs) in wastewaters. The present work aimed to build on the current knowledge about the effects of activated sludge and UV irradiation on antibiotic resistance determinants in biologically treated wastewaters. For that, the microbial community and ARGs' composition of samples collected after preliminary (APT), secondary (AST), and tertiary (ATT) treatments in a full-scale wastewater treatment plant using a modified activated sludge (MAS) system followed by an UV stage (16 mJ/cm(2)) were investigated through culture-dependent and independent approaches (including metagenomics). A total of 24 phyla and 460 genera were identified, with predominance of Gammaproteobacteria in all samples. Pathogenic genera corresponded to 8.6% of all sequences on average, mainly Acinetobacter and Streptococcus. Significant differences (p < 0.05) in the proportion of pathogens were observed between APT and the other samples, suggesting that the secondary treatment reduced its abundance. The MAS achieved 64.0-99.7% average removal efficiency for total (THB) and resistant heterotrophic bacteria, although the proportions of ARB/THB have increased for sulfamethoxazole, cephalexin, ciprofloxacin, and tetracycline. A total of 10(7) copies/mL of intI1 gene remained in the final effluent, suggesting that the treatment did not significantly remove this gene and possibly other ARGs. In accordance, metagenomic results suggested that number of reads recruited to plasmid-associated ARGs became more abundant in the pool throughout the treatment, suggesting that it affected more the bacteria without these ARGs than those with it. In conclusion, disinfected effluents are still a potential source for ARB and ARGs, which highlights the importance to investigate ways to mitigate their release into the environment.202235060061
7202170.9998Cyanobacterial extracellular antibacterial substances could promote the spread of antibiotic resistance: impacts and reasons. Many studies have shown that antibiotic resistance genes (ARGs) can be facilitated by a variety of antibacterial substances. Cyanobacteria are photosynthetic bacteria that are widely distributed in the ocean. Some extracellular substances produced by marine cyanobacteria have been found to possess antibacterial activity. However, the impact of these extracellular substances on ARGs is unclear. Therefore, we established groups of seawater microcosms that contained different concentrations (1000, 100, 10, 1, 0.1, 0.01, and 0 μg mL(-1)) of cyanobacterial extracellular substances (CES), and tracked the changes of 17 types of ARGs, the integron gene (intI1), as well as the bacterial community at different time points. The results showed that CES could enrich most ARGs (15/17) in the initial stage, particularly at low concentrations (10 and 100 μg mL(-1)). The correlation analysis showed a positive correlation between several ARGs and intI1. It is suggested that the abundance of intI1 increased with CES may contribute to the changes of these ARGs, and co-resistance of CES may be the underlying reason for the similar variation pattern of some ARGs. Moreover, the results of qPCR and high-throughput sequencing of 16S rRNA showed that CES had an inhibitory impact on the growth of bacterial communities. High concentrations of CES were found to alter the structure of bacterial communities. Co-occurrence networks showed that bacteria elevated in the high concentration group of CES and might serve as the potential hosts for a variety of ARGs. In general, marine cyanobacteria could play an important role in the global dissemination of ARGs and antibiotic-resistant bacteria (ARBs).202337947439
7845180.9998Mechanism and potential risk of antibiotic resistant bacteria carrying last resort antibiotic resistance genes under electrochemical treatment. The significant rise in the number of antibiotic resistance genes (ARGs) that resulted from our abuse of antibiotics could do severe harm to public health as well as to the environment. We investigated removal efficiency and removal mechanism of electrochemical (EC) treatment based on 6 different bacteria isolated from hospital wastewater carrying 3 last resort ARGs including NDM-1, mcr-1 and tetX respectively. We found that the removal efficiency of ARGs increased with the increase of both voltage and electrolysis time while the maximum removal efficiency can reach 90%. The optimal treatment voltage and treatment time were 3 V and 120 min, respectively. Temperature, pH and other factors had little influence on the EC treatment process. The mechanism of EC treatment was explored from the macroscopic and microscopic levels by scanning electron microscopy (SEM) and flow cytometry. Our results showed that EC treatment significantly changed the permeability of cell membrane and caused cells successively experience early cell apoptosis, late cell apoptosis and cell necrosis. Moreover, compared with traditional disinfection methods, EC treatment had less potential risks. The conjugative transfer frequencies of cells were significantly reduced after treatment. Less than 1% of bacteria entered the viable but nonculturable (VBNC) state and less than 5% of intracellular ARGs (iARGs) turned into extracellular ARGs (eARGs). Our findings provide new insights into as well as important reference for future electrochemical treatment in removing ARB from hospital wastewater.202235085630
7966190.9998How heavy metal stress promotes dissemination of antibiotic resistance genes in the activated sludge process. Heavy metals have been recently revealed as promoters to antibiotic resistance gene (ARG) dissemination in water environment, but their influence on ARG transfer in the activated sludge process has not been clear. In this study, a set of sequencing batch reactors (SBRs) and micro-scale microfluidic chips were established to quantify the impacts of heavy metals (0.5 mM of Pb, 0.1 mM of As, and 0.005 mM of Hg) on the ARG spreading in the activated sludge consortium. Under heavy metal stress, transfer frequencies were 1.7-3.6 folds increase compared to the control. Gram-negative bacteria increased significantly after heavy metal added, which were more prone to receiving resistant plasmid from donors. Meanwhile, the relative expression of genes related to conjugation changed in activated sludge, especially the expression of outer membrane protein and oxidative stress regulatory genes increased by 2.9-7.4 folds and 7.8-13.1 folds, respectively. Furthermore, using microfluidic chips, the dynamics of ARG transfer was observed at single cell level under heavy metal pressure. Heavy metals firstly promoted conjugation and then vertical gene transfer played an important part for ARG spreading. The results provided in-depth understanding of the influence of heavy metals on ARG behavior in the environment.202235724617